Single-Use Tangential Flow Filtration for Closed Processing
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The document discusses the single-use tangential flow filtration (TFF) systems offered by Merck KGaA, specifically focusing on closed processing to minimize contamination risks and improve biopharmaceutical manufacturing efficiency. It outlines definitions, advantages, and the evolution toward fully closed systems, detailing components, processes, and performance testing of the TFF systems. The overall aim is to assure product integrity while facilitating faster production and reducing reliance on cleanroom environments.
![Introduction to Equipment used in Experimental Set-up
Mobius® FlexReady System for TFF and Pellicon® Capsule
Mobius® FlexReady
System for TFF
Pellicon® Capsule
& Manifold
Available Recycle Tank Volume [L] 50, 100, 200 -
Membrane Area [m2] 0.5 – 5
0.5 m² - 1.5 m²
Manifold: 1 m²
Multiple of 1.5m²
Flowrate [L/min]
2 – 20
Reco. flowrates for TFF
4-8 L/min/m2
-
Details
Pre-sterilized flowpath
sub-assemblies
Pre-sterilized
ready-to-connect
17](https://image.slidesharecdn.com/closedtffprocessinginmobiustf2ssystemwebinarcontent-fnl002-210507112938/85/Single-Use-Tangential-Flow-Filtration-for-Closed-Processing-17-320.jpg)
![Low Viscosity Applications, Manual Mode of Operation
25
Product Recovery
Protein Recovery Summary
Conc, Pre-Recovery [g/L] 53
Viscosity [cP] 1.6
Yield [%] 102.9
Final Conc [g/L] 47
Step 3 Step 4
Step
1&
2](https://image.slidesharecdn.com/closedtffprocessinginmobiustf2ssystemwebinarcontent-fnl002-210507112938/85/Single-Use-Tangential-Flow-Filtration-for-Closed-Processing-25-320.jpg)
![Elevated Viscosity Applications, Manual Mode of Operation
26
Product Recovery
Protein Recovery Summary
Conc, Pre-Recovery [g/L] 186
Viscosity [cP] 10
Yield [%] 99.0
Final Conc [g/L] 165
Step 3 Step 4
Step
1&
2](https://image.slidesharecdn.com/closedtffprocessinginmobiustf2ssystemwebinarcontent-fnl002-210507112938/85/Single-Use-Tangential-Flow-Filtration-for-Closed-Processing-26-320.jpg)
![Low Viscosity Applications, Automated Mode of Operation
28
Product Recovery
Protein Recovery Summary
Conc, Pre-Recovery [g/L] 51
Viscosity [cP] 1.5
Yield [%] 97.6
Final Conc [g/L] 44
Step 3 Step 4
Step 1& 2](https://image.slidesharecdn.com/closedtffprocessinginmobiustf2ssystemwebinarcontent-fnl002-210507112938/85/Single-Use-Tangential-Flow-Filtration-for-Closed-Processing-28-320.jpg)
![Elevated Viscosity Applications, Automated Mode of Operation
29
Product Recovery
Protein Recovery Summary
Conc, Pre-Recovery [g/L] 202
Viscosity [cP] 14.5
Yield [%] 94.9
Final Conc [g/L] 170
Step 3 Step 4
Step 1& 2](https://image.slidesharecdn.com/closedtffprocessinginmobiustf2ssystemwebinarcontent-fnl002-210507112938/85/Single-Use-Tangential-Flow-Filtration-for-Closed-Processing-29-320.jpg)
Single-Use Tangential Flow Filtration for Closed Processing
- 1. The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada. Single-Use Tangential Flow Filtration for Closed Processing May 6, 2021
- 2. The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada. Sarah Le Merdy Strategy Deployment, Single-Use and Integrated Systems Dana Kinzlmaier Applications Engineer SPEAKERS:
- 3. The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada
- 4. Agenda 1 2 3 Definitions and drivers TFF processing in a closed mode System characterization and performance testing 4 Conclusion and perspectives
- 6. 6 What is closed processing about? Isolating a product from the environment and vice versa “A process that ensures that biological material does not come into contact with the external environment. Closed processing enables grey space or CNC (Controlled Not Classified) processing and is sometimes referred to as being conducted in a “ballroom”. According to ISPE: “A process condition when the product, materials, critical components or container/closure surfaces are contained and separated from the immediate process environment within closed/sealed process equipment. A process step (or system) in which the product and product contact surfaces are not exposed to the immediate room environment.” BPOG: “The key factor for industry acceptance was the Pharmaceutical Inspection Cooperation Scheme (PIC/S) determination that clean-room requirements for bioP can be relaxed/eliminated if your process is closed”. ISPE Baseline Guide: “The strategy of using uncontrolled or unclassified spaces for closed piping systems has always been accepted by the regulatory agencies as an acceptable risk”.
- 7. 7 FUNCTIONALLY CLOSED FULLY CLOSED BRIEFLY OPEN SYSTEM • Never exposed to the environment • Materials may be introduced to a closed system, in a way to avoid exposure of the product to the room environment • Exposes the product to the room environment • The room environment is controlled to minimize the risk of product contamination • For bioburden-controlled processing, open operations are expected to be performed in a classified environment • May be routinely opened but returned to a closed state through a sanitization or sterilization step prior to process use • Defining and validating the required sanitization or sterilization is the user's responsibility Source : Challenging the Cleanroom Paradigm for Biopharmaceutical Manufacturing of Bulk Drug Substances Aug 01, 2011, http://www.biopharminternational.com/challenging-cleanroom-paradigm-biopharmaceutical-manufacturing-bulk-drug-substances?id=&sk=&date=&pageID=2 … and many different users' interpretations! How closed? Different ways of achieving a closed state
- 8. Drivers and benefits ‘Closed’ single-use is an accelerator of single-use benefits 1 Protect from contamination – Ultimately reducing cleanroom requirements by minimizing contamination independently from the production environment 2 Flexibility to market - Means to reduce time to market using multimodal solutions and/or reducing turnover time between campaigns, in grey space 3 Reduction of interferences – Limiting the number of human manipulations and high-risk open state throughout the process to limit risk of failure 8
- 9. Early and late steps are typically closed by definition Cell culture, upstream and final fill: - Closed by design - Closed by operation Downstream culture of reuse - Reusable expendables (TFF devices, chromatography media) with optimized lifecycle - Steps still to be converted to single-use operation (clarification) or with large footprint Drivers and benefits Closing what is currently open: Downstream steps 9 Moving downstream steps to single-use and ultimately closed technologies is a natural next step of the evolution of biomanufacturing
- 10. Drivers and benefits Different drivers lead to closed processing 10 • Manufacturing at ‘point of use’ • Small, fast set-up capabilities • Safety of product and patient “The main objective is the assurance of an uncompromised product using a processing mode which doesn't have potentially harmful steps.” • Reduce human interferences • Reduce environment control requirements “The flowpaths used in closed mode help increasing process safety and participate to easier, faster, and lighter environmental control.” • Filtration not possible • Biosafety: no egress • Animal vaccine never sterile filtered Future state: • Multimodal solutions • Ballroom concept Viral vector & live viruses Recombinant proteins Cell therapy
- 11. TFF Processing in closed mode
- 12. Open versus closed processing step Tighter control and management of inputs and outputs in closed Open Multi-use sensors can be used Ideally fully automated ‘All’ expendable Similar to MU processing ‘Open’ lines authorized, connect/disconnect containers Similar to multi-use TFF Similar to multi- use TFF Typically class C SYSTEM & FLOWPATH EXPENDABLE SET-UP PROCESSING SAMPLING ENVIRONMENT Closed (Fct) SU sensing technologies Aseptic Connectors Ideally fully auto Can be sanitized or sterilized in place Collecting bags for all fluids from sanitization step Post-sanitization all fluids in/out are contained Closed Potentially lower to class D Open allowed post- product recovery Closed (Fully) Drip-less disconnections Are pre-sterilized and equipped with aseptic connectors or built in flowpath Bags for all fluids from process start (water, buffers, waste/drain, product…) From set-up to dismounting all fluids are contained Closed, ideally part of the flowpath assemblies Potentially lower to class D Potentially ‘never’ open, closed and dripless disconnections/ dismounting/ discarding 12
- 13. Open: Connections and disconnections in a non-sterile environment during the different steps in the TFF operation (flushing, cleaning, draining, etc) Flowpath and system design Closed operation flowpath requires additional fixed lines WFI Air NaOH Buffer Product 13 Filtrate
- 14. Flowpath and system design Closed operation flowpath requires additional fixed lines Closed: Assemblies used for cleaning, flushing and draining incorporated into one single assembly (multiple sub-assemblies mated with aseptic connectors) Control of fluid flow during flushing and draining done with pneumatic pinch clamps 14 Filtrate WFI Air NaOH Buffer Product
- 15. Functionally closed - Select any device format which can be sanitized in place - Take into account sanitization agent and flushing volumes Expendable selection 15 Fully closed - Select pre-sterilized device format - Take into account connectivity/adaptability to flowpath Closed-enabled formats It is not necessarily about selecting a ‘new’ or ‘specific’ filtration device type…
- 16. System Characterization and Performance Testing Flexware® Assemblies for Closed TFF Processing
- 17. Introduction to Equipment used in Experimental Set-up Mobius® FlexReady System for TFF and Pellicon® Capsule Mobius® FlexReady System for TFF Pellicon® Capsule & Manifold Available Recycle Tank Volume [L] 50, 100, 200 - Membrane Area [m2] 0.5 – 5 0.5 m² - 1.5 m² Manifold: 1 m² Multiple of 1.5m² Flowrate [L/min] 2 – 20 Reco. flowrates for TFF 4-8 L/min/m2 - Details Pre-sterilized flowpath sub-assemblies Pre-sterilized ready-to-connect 17
- 18. Mobius® FlexReady System for TFF Fully Closed Flowpath 1 Recycle Vessel Assembly 100 L 2 Transfer Pump Assembly 3 Feed Pump Assembly 4 Transfer Pump Manifold Assembly 5 Smart Flexware® Assembly 6 Pellicon® Capsule Assembly 7 Retentate Sampling Port Assembly 8 Flush/Recovery Assembly 9 Permeate Assembly (with SU sensor flow cell) 10 Permeate Sampling Port Assembly 1 4 2 3 7 5 8 9 10 6 18
- 19. 19 Application Testing System Characterization and Process Testing 1 Minimum working volume As a function of flow rate and viscosity 2 Flowpath pressure drop As a function of viscosity Total pressure drop Retentate line pressure drop 3 Process run Concentration and Diafiltration of Human Plasma IgG 4 Product recovery Low viscosity applications Elevated viscosity applications
- 20. System Characterization Minimum Working Volume 20 Is the minimum volume at which the system can be operated at given process flowrate without causing air entrainment in the process fluid. It accounts for volume in the tubing, valves, instruments, membranes, and minimal volume in the recycle tank. Why is it important? To ensure air is not entrained into the system during processing. Minimum Working Volume Minimum working volume of flowpath with 1 m2 Pellicon® Capsule assembly at a typical feed flow rate of 4 - 8 L/min is <1.3 L for solutions with viscosity between 1 – 50 cP • Capsule assembly not installed during testing • Measurements performed with • Purified Water: 1 cP viscosity • Glycerin/Purified Water solution: • 10, 25 and 50 cP viscosity
- 21. System Characterization Total Pressure Drop of the System 21 Total pressure drop of a system is the line drop from the feed pump discharge through the feed lines, devices and retentate line back to the recycle vessel. Why is it important? High Total System Pressure Drop could limit the flow rate that can be driven through devices to increase mass transfer and drive high flux. It is especially true for high protein concentrations and high viscosity solutions. • Capsule assembly not installed during testing • Measurements performed with purified water and Mixture of Glycerin/ Purified Water Total Pressure Drop of the system in the flow rate range of 4-8 L/min is less than 6 psi in 1- 50 cP viscosity range No significant pressure drop increase due to added sterile connectors
- 22. System Characterization Retentate Pressure Drop 22 The retentate pressure drop is the line drop from the outlet of the membrane holder through the retentate line back to the recycle vessel. Why is it important? High retentate line pressure drop could limit the minimum transmembrane pressure that can be achieved even with the retentate control valve fully open. • Capsule assembly not installed during testing • Measurements performed with Purified Water and Mixture of Glycerin/ Purified Water Retentate Pressure Drop in the flow rate range of 4-8 L/min is less than 5 psi in 1- 50 cP viscosity range No significant pressure drop increase due to added sterile connectors
- 23. Equilibration Concentration Diafiltration Final concentration Recovery Closed Processing with Single Use Flexware® Assemblies and Devices 23 • Equilibration buffer 10 mM PBS, pH 7.2 • 20 L/m2 18.5 L 20 g/L to 60 g/L IgG Viscosity: 1.5 – 1.8 cP Diafiltration at 6 L 25 mM Sodium Acetate, pH 5.5 8 Diavolumes 60 g/L to 200 g/L IgG Viscosity: 1.8 – 13 cP Elevated Viscosity Applications Low Viscosity Applications Manual & Automated All streams have to be collected into bags Buffer Product Waste Permeate Product In Waste Retentate Out
- 24. Product Recovery XV001 XV009 XV003 XV002 XV404 XV102 PCV101 Recycle Vessel XV441 XV402 XV431 XV421 XV401 XV411 XV403 XV405 XV007 XV006 XV406 WFI F401 NaOH Buffer Source Drain Recovery Drain To Flush Bag P002 P001 XV004 XV005 PCV001 XV001 XV009 XV003 XV002 XV404 XV102 PCV101 Recycle Vessel XV441 XV402 XV431 XV421 XV401 XV411 XV403 XV405 XV007 XV006 XV406 WFI F401 NaOH Buffer Source Drain Recovery Drain To Flush Bag P002 P001 XV004 XV005 PCV001 XV001 XV009 XV003 XV002 XV404 XV102 PCV101 Recycle Vessel XV441 XV402 XV431 XV421 XV401 XV411 XV403 XV405 XV007 XV006 XV406 WFI F401 NaOH Buffer Source Drain Recovery Drain To Flush Bag P002 P001 XV004 XV005 PCV001 XV001 XV009 XV003 XV002 XV404 XV102 PCV101 Recycle Vessel XV441 XV402 XV431 XV421 XV401 XV411 XV403 XV405 XV007 XV006 XV406 WFI F401 NaOH Buffer Source Drain Recovery Drain To Flush Bag P002 P001 XV004 XV005 PCV001 24 Step 1 Pump the Product out of the recycle vessel through the recovery line Step 2 Gravity Drain the Retentate Line between the bottom of the recycle vessel and transfer line Step 3 Buffer Flush through the transfer and retentate lines, devices to the recovery line Step 4 Flush the Retentate Line between the transfer line and bottom of the recycle vessel with a small amount of buffer, then pump it out through the recovery line
- 25. Low Viscosity Applications, Manual Mode of Operation 25 Product Recovery Protein Recovery Summary Conc, Pre-Recovery [g/L] 53 Viscosity [cP] 1.6 Yield [%] 102.9 Final Conc [g/L] 47 Step 3 Step 4 Step 1& 2
- 26. Elevated Viscosity Applications, Manual Mode of Operation 26 Product Recovery Protein Recovery Summary Conc, Pre-Recovery [g/L] 186 Viscosity [cP] 10 Yield [%] 99.0 Final Conc [g/L] 165 Step 3 Step 4 Step 1& 2
- 27. 27 Product Recovery Automation: A Key to Process Consistency Common Control Platform® (CCP®) Software • Written application recipes can be edited and stored for different processes • Report generator quickly creates batch reports Advantages of Automation • Run to run consistency • Lower risk of operator error • Easier troubleshooting Product Recovery, Automated Mode Product recovery recipes written, tested and optimized to achieve maximum product recovery with minimal dilution.
- 28. Low Viscosity Applications, Automated Mode of Operation 28 Product Recovery Protein Recovery Summary Conc, Pre-Recovery [g/L] 51 Viscosity [cP] 1.5 Yield [%] 97.6 Final Conc [g/L] 44 Step 3 Step 4 Step 1& 2
- 29. Elevated Viscosity Applications, Automated Mode of Operation 29 Product Recovery Protein Recovery Summary Conc, Pre-Recovery [g/L] 202 Viscosity [cP] 14.5 Yield [%] 94.9 Final Conc [g/L] 170 Step 3 Step 4 Step 1& 2
- 30. 30 Summary Closed TFF System and Process Characterization 1 Minimum working volume • Minimum working volume using 1 m² Pellicon® Capsule assembly is < 1.3 L for solutions with viscosity between 1 – 50 cP 2 Flowpath pressure drop • No significant pressure drop increase due to added sterile connectors • For typical range of flow rates for a TFF process and viscosity 1-50 cp • Total system: < 6 psi • Retentate line: < 5 psi 3 Process run • Protein solutions can be concentrated to high final concentrations • System can handle processing elevated viscosity solutions with ease 4 Product recovery • Proposed recovery methods result in ≥ 95% protein recoveries with minimal protein dilution • Process can be adjusted to comply with customer specifications and needs
- 31. 31 A closed state can be achieved in different ways and would present benefits for downstream steps. While drivers vary between applications, the same specific mindset needs applying to make it successful with no compromise on performance. Ultimately, closed processing steps will be an integral part of connected, continuous processing. Conclusion and perspectives
- 32. sarah.lemerdy@emdgroup.com dana.kinzlmaier@milliporesigma.com Sarah Le Merdy Dana kinzlmaier The vibrant M, Millipore, Flexware, Mobius and Pellicon are trademarks of Merck KGaA, Darmstadt, Germany or its affiliates. All other trademarks are the property of their respective owners. Detailed information on trademarks is available via publicly accessible resources. © 2021 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved. Acknowledgments Special Thanks to North America MSAT Team, Patrick McGee, Elizabeth Goodrich, Akshat Gupta, Emily Peterson, Gayathri Raja and Product Marketing Team, Sylvie Riou and Christophe Dufossé Thank you